New composition

ABSTRACT

Compositions and formulations comprising a non-neutralised tocol phosphate and a vitamin A compound, which are suitable for the treatment of inflammation and/or infection in breast or udder tissue, more particularly in a mammary gland, reducing the somatic cell count in a lactating subject and supplementing vitamin E levels in a subject.

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application is a continuation of U.S. patent applicationSer. No. 14/004,973, filed Sep. 13, 2013, which is a national stagefiling under 35 U.S.C. §371 of International Application No.PCT/AU2012/000220, filed on 2 Mar. 2012, which claims the benefit ofU.S. Provisional Application No. 61/452,692, filed 15 Mar. 2011, andforeign priority to Australian Patent Application No. 2011902893, filedon 20 Jul. 2011. Priority to each application is hereby claimed.

TECHNICAL FIELD

The present invention relates to new compositions suitable for thetreatment of inflammation and/or infection in breast or udder tissue,more particularly in a mammary gland. The inflammation and/or infectionin a mammary gland may be mastitis.

BACKGROUND

Mastitis is associated with inflammation and/or infection in breast orudder tissue, more particularly in a mammary gland.

Mastitis is a serious problem in all mammals, including humans andanimals.

Mastitis in dairy cattle, for example, is an economically devastatingdisease causing immense economic losses in the dairy industry and is themost expensive production disease in dairy herds worldwide. For example,in Australia and New Zealand combined, there are about 6 million cattleproducing milk. About 15% of any given herd suffers clinical orsub-clinical mastitis, and therefore, at any given time there are about900,000 cattle in Australia and New Zealand alone that are sufferingfrom clinical and sub-clinical mastitis. Currently, every case ofclinical mastitis costs farmers at least $200.

The sub-clinical or clinical definition of mastitis is defined by thepresence of somatic cells in milk. In Australia and other countries, thepayment to the farmer for the milk may be somewhat dependent on somaticcell count. For example, at very low somatic cell counts a premium pricemay be paid for the milk. On the other hand, milk above a thresholdsomatic cell count of 400,000 cells/ml is not considered fit for humanconsumption in Europe.

Current treatments for mastitis include long-acting antibiotics, such asampicillin, cloxacilin, cephalonium and bismuth subnitrate.

With respect to dairy cattle, in addition to the costs of administeringthe antibiotics, there is also is a withholding period that must elapsebefore the milk produced by treated subjects can be used for humanconsumption. This withholding period varies between the types ofantibiotics administered, but is typically about 96 hours. Thiswithholding period results in significant losses in productivity fordairy farms.

With respect to humans, antibiotic treatment may affect breastfeeding.

Accordingly, there is a need for improved or alternative treatments forinflammation and/or infection in breast or udder tissue, moreparticularly in a mammary gland, for use in livestock, but also in othermammals such as humans, and to address the problems caused by mastitis.

In this specification where a document, act or item of knowledge isreferred to or discussed, this reference or discussion is not anadmission that the document, act or item of knowledge or any combinationthereof was at the priority date, publicly available, known to thepublic, part of common general knowledge; or known to be relevant to anattempt to solve any problem with which this specification is concerned.

SUMMARY

The present invention relates to compositions comprising anon-neutralised tocol phosphate and a vitamin A compound. It has beenfound that the compositions are suitable for the treatment ofinflammation and/or infection in breast or udder tissue, moreparticularly is in a mammary gland. The inflammation and/or infection ina mammary gland may be mastitis. It would advantageous if the presentinvention avoids the need for antibiotic treatments.

Accordingly, a first aspect of the present invention provides acomposition comprising a non-neutralised tocol phosphate and a vitamin Acompound.

The non-neutralised tocol phosphate may be a non-neutralised tocopherylphosphate, a non-neutralised tocotrienyl phosphate, or a combinationthereof.

The non-neutralised tocol phosphate may be selected from the groupconsisting of non-neutralised mono-(tocopheryl) phosphate,non-neutralised mono-(tocopheryl) phosphate monosodium salt,non-neutralised mono-(tocopheryl) phosphate disodium salt,non-neutralised mono-(tocopheryl) phosphate monopotassium salt,non-neutralised mono-(tocopheryl) phosphate dipotassium salt,non-neutralised di-(tocopheryl) phosphate, non-neutraliseddi-(tocopheryl) phosphate monosodium salt, non-neutraliseddi-(tocopheryl) phosphate monopotassium salt, non-neutralisedmono-(tocotrienyl) phosphate, non-neutralised mono-(tocotrienyl)phosphate monosodium salt, non-neutralised mono-(tocotrienyl) phosphatedisodium salt, non-neutralised mono-(tocopheryl) phosphate monopotassiumsalt, non-neutralised mono-(tocotrienyl) phosphate dipotassium salt,non-neutralised di-(tocotrienyl) phosphate, non-neutraliseddi-(tocotrienyl) phosphate monosodium salt, non-neutraliseddi-(tocotrienyl) phosphate monopotassium salt, or a combination thereof.

In some embodiments, the non-neutralised tocol phosphate is acombination of a non-neutralised mono-(tocopheryl) phosphate,non-neutralised mono-(tocopheryl) phosphate monosodium salt,non-neutralised mono-(tocopheryl) phosphate disodium salt,non-neutralised mono-(tocopheryl) phosphate monopotassium salt ornon-neutralised mono-(tocopheryl) phosphate dipotassium salt and anon-neutralised di-(tocopheryl) phosphate, non-neutraliseddi-(tocopheryl) phosphate monosodium salt or non-neutraliseddi-(tocopheryl) phosphate monopotassium salt. In other embodiments, thenon-neutralised tocol phosphate is a combination of a non-neutralisedmono-(tocotrienyl) phosphate, non-neutralised mono-(tocotrienyl)phosphate monosodium salt, non-neutralised mono-(tocotrienyl) phosphatedisodium salt, non-neutralised mono-(tocopheryl) phosphate monopotassiumsalt or non-neutralised mono-(tocotrienyl) phosphate dipotassium saltand a non-neutralised di-(tocotrienyl) phosphate, non-neutraliseddi-(tocotrienyl) phosphate monosodium salt or non-neutraliseddi-(tocotrienyl) phosphate monopotassium salt.

In one embodiment, the combination is a non-neutralisedmono-(tocopheryl) phosphate and a non-neutralised di-(tocopheryl)phosphate.

When the composition comprises a combination, the ratio (w/w %) ofnon-neutralised mono-(tocol) phosphate to non-neutralised di-(tocol)phosphate may be at least 2:1, or about 4:1 to about 1:4, or about 6:4to about 8:2. In some embodiments, the ratio may be about 2:1, about6:4, or about 8:2.

The pH of the non-neutralised tocol phosphate may be less than about 4.In some embodiments, the pH of the non-neutralised tocol phosphate isabout 2 to about 4, or about 2 to about 3. In particular embodiments,the pH of the non-neutralised tocol phosphate is about 2 or about 3.

The non-neutralised tocol phosphate may comprise from about 0.01% w/w upto about 40% w/w, from about 0.01% w/w up to about 30% w/w, from about0.01% w/w up to about 20% w/w, from about 0.01% w/w up to about 10% w/w,from about 0.01% w/w up to about 5% w/w, from about 0.1% w/w up to about5% w/w, from about 0.1% w/w up to about 2.5% w/w, from about 0.1% w/w upto about 0.5% w/w, or about 0.5% w/w, about 1% w/w, about 2% w/w, about3% w/w, about 5% w/w, about 10% w/w, or about 20% w/w, of the totalconcentration of the composition.

The vitamin A compound may be selected from the group consisting ofvitamin A, a derivative of vitamin A, a metabolite of vitamin A, aprecursor of vitamin A, or pro-vitamin A, or a combination thereof.

The vitamin A compound may be a retinoid. The retinoid may be selectedfrom the group consisting of retinol, retinal (retinaldehyde), tretinoin(retinoic acid), isotretinoin, alitretinoin, etretinate and itsmetabolite acitrein, tazarotene, bexarotene and adapalene. The vitamin Acompound may also be a retinyl ester such as retinyl acetate or retinylpalmitate.

In one embodiment, the vitamin A compound is a carotenoid. Thecarotenoid may be of the xanthophylls class such as lutein andzeaxanthin or the carotene class such as alpha-carotene, beta-carotene,gamma-carotene, beta-cryptoxanthin and lycopene. In a preferredembodiment, the vitamin A compound is beta-carotene.

In some embodiments, the composition may comprise a combination of amono-(tocopheryl) phosphate and a di-(tocopheryl) phosphate, andbeta-carotene.

The ratio (w/w %) of non-neutralised tocol phosphate to vitamin Acompound may be from about 0.01:1 to about 100:1, from about 0.01:1 toabout 5:1, from about 0.05:1 to about 2:1, from about 1:1 to about 5:1,from about 10:1 to about 60:1, or from about 30:1 to about 50:1. In someembodiments, the ratio may be about 0.05:1, about 0.1:1, about 1:1,about 5:1, about 10:1, about 25:1, or about 40:1.

The composition may optionally comprise a delivery vehicle. Accordingly,a second aspect of the present invention provides a formulationcomprising the composition as defined above and a delivery vehicle. In apreferred embodiment, the delivery vehicle is a hydrophobic deliveryvehicle. The hydrophobic delivery vehicle may be a solid or a liquid.The hydrophobic delivery vehicle may be an oil or a wax. Preferably, thehydrophobic delivery vehicle is an oil such as olive oil.

The delivery vehicle may be present in an amount of at least about 60.0%w/w, at least about 80.0% w/w, at least about 90% w/w, at least about99.0% w/w, or at least about 99.5% w/w, of the total concentration ofthe formulation. In some embodiments, the delivery vehicle is present inan amount of about 60.0% w/w, about 80.0% w/w, about 90% w/w, about 98%w/w, about 99% w/w, or about 99.5% w/w, of the total concentration ofthe formulation.

The compositions and the formulations of the present invention mayoptionally comprise a nutrient compound. The nutrient compound may beselected from the group consisting of coenzyme Q10 (or ubiquinone);ubiquinol; fat-soluble vitamins such as vitamin

D compounds (e.g. D2, D3 and their derivatives), vitamin E compounds (α,β, γ, δ-tocopherols, or, α, β, γ, δ-tocotrienols), vitamin K compounds(e.g. K1, K2, K3 and their derivatives); folic acid; vitamin B compounds(e.g. B1, B2, B3, B6, and B12); vitamin C; vitamin P; vitamin F; lutein;zeaxanthin; cysteine; flavonoids (e.g. catechins, reserveratrol,proanthocyanidins); isoflavones (e.g. genistein and daidzein); bilberry;ginkgo biloba; grape seed extract; phytonutrients (e.g. lycopene, luteinand seaxanthin); alpha lipoic acid; bilberry; bioflavinoids; unsaturatedfatty acids (e.g. linoleic acid, conjugated linoleic acid, linolenicacid, omega-3 fatty acids such as docosahexaenoic acid (DHA) andeicosapentaeonic acid (EPA) and their glycerol-esters); calcium;phosphorus; magnesium; fluorine; phosphorus; sulfur; sodium; potassium;chloride; calcium; iodine; cobalt; copper; iron; manganese; molybdenum;selenium; zinc; chromium; cadmium; fluorine; nickel; silicon; tin;vanadium; niacin; and combinations thereof.

The compositions and the formulations of the present invention may beprepared by a variety of techniques. Accordingly, a third aspect of thepresent invention provides methods for preparing the composition and theformulation as defined above.

One method of preparing the composition defined above comprises thesteps of mixing a non-neutralised tocol phosphate and a vitamin Acompound, in suitable quantities, with stirring, until completehomogenisation is achieved. Another method of preparing the compositiondefined above comprises the steps of warming the non-neutralised tocolphosphate to a temperature greater than about 60° C., preferably betweenabout 70° C. and about 80° C., and then adding the vitamin A compound tothe non-neutralised tocol phosphate when the non-neutralised tocolphosphate is at a temperature of less than about 40° C., with stirring,until complete homogenisation is achieved. One method for preparing theformulation as defined above comprises the steps of mixing thenon-neutralised tocol phosphate and the hydrophobic delivery vehicle,and warming the mixture to a temperature greater than about 60° C.,preferably between about 70° C. and about 80° C., and then adding thevitamin A compound to the non-neutralised tocol phosphate when thenon-neutralised tocol phosphate is at a temperature of less than about40° C., with stirring, until complete homogenisation is achieved.

The compositions and the formulations of the present invention aresuitable for the treatment of inflammation and/or infection in breast orudder tissue, more particularly in a mammary gland. The inflammationand/or infection in a mammary gland may be mastitis.

Accordingly, a fourth aspect of the present invention provides a methodfor the treatment of inflammation and/or infection in breast or uddertissue comprising administering the composition or the formulation asdefined above to a subject in need thereof. The present invention alsoprovides use of a non-neutralised tocol phosphate and a vitamin Acompound for the treatment of inflammation and/or infection in breast orudder tissue. The zo present invention further provides use of anon-neutralised tocol phosphate and a vitamin A compound in themanufacture of a medicament for the treatment of inflammation and/orinfection in breast or udder tissue.

The breast or udder tissue may be a mammary gland.

The inflammation and/or infection in the mammary gland may be mastitis,more particularly puerperal mastitis.

The subject may be a mammal. The mammal may be selected from humans,domestic mammals such as companion animals, working animals, livestock,and zoological/park mammals. Preferably, the subject is a mammal, inparticular a female mammal, more particularly a female mammal havingdeveloped lactiferous ducts such as a lactating female mammal. Examplesof female lactating mammals may be humans or ungulates (even-toed andodd-toed) including cows, goats, sheep, yaks, water buffalos, horses,reindeer, camels, alpacas, bantengs, donkeys, oxen, zebu, moose andbison.

The compositions or the formulations may be administered topically,preferably epicutaneus, or enterally, preferably orally.

Dosage forms for topically or enteral administration are also preferred.The compositions or the formulations may be administered weekly, daily,or multiple times per day.

The dosage level of the non-neutralised tocol phosphate may be about 0.1to about 20 mg per kg subject body weight per day which can beadministered in single or multiple doses. Preferably, the dosage levelwill be about 0.1 to about 15 mg/kg per day; about 0.1 to about 10 mg/kgper day; more preferably, about 0.1 to about 10 mg/kg per day, about 0.1to about 5 mg/kg per day, about 0.1 to about 2.5 mg/kg per day, or about0.1 to about 2 mg/kg per day. A suitable dosage level may be about 0.1to about 7.5 mg/kg per day. For example, within the above dosage ranges,the dosage may be about 0.1 mg/kg per day, about 0.2 mg/kg per day,about 0.4 mg/kg per day, about 0.6 mg/kg per day, about 0.7 mg/kg perday, about 0.8 mg/kg per day, about 0.9 mg/kg per day, about 1 mg/kg perday, about 1.2 mg/kg per day, about 1.4 mg/kg per day, about 1.5 mg/kgper day, about 1.6 mg/kg per day, about 1.7 mg/kg per day, about 1.8mg/kg per day, about 1.9 mg/kg per day, about 2 mg/kg per day, about 2.2mg/kg per day, about 2.5 mg/kg per day, about 3 mg/kg per day, about 5mg/kg per day, about 7.5 mg/kg per day, or about 10 mg/kg per day.

The dosage level of the vitamin A compound may be about 10 to about 1000μg per kg subject body weight per day which can be administered insingle or multiple doses. Preferably, the dosage level will be about 10to about 850 μg/kg per day; about 10 to about 800 μg/kg per day; about10 to about 500 μg/kg per day; or less than about 300 μg/kg per day.More preferably, the dosage level will be about 10 to about 500 μg/kgper day, 10 to about 275 μg/kg per day, 10 to about 250 μg/kg per day,20 to about 250 μg/kg per day, 25 to about 250 μg/kg per day, 10 toabout 200 μg/kg per day, or 50 to about 200 μg/kg per day. A suitabledosage level may be about 10 to about 300 μg/kg per day. For example,within the above dosage ranges, the dosage may be about 10 μg/kg perday, about 20 μg/kg per day, about 40 μg/kg per day, about 50 μg/kg perday, about 60 μg/kg per day, about 70 μg/kg per day, about 75 μg/kg perday, about 80 μg/kg per day, about 85 μg/kg per day, about 90 μg/kg perday, about 95 μg/kg per day, about 100 μg/kg per day, about 125 μg/kgper day, about 150 μg/kg per day, about 155 μg/kg per day, about 160μg/kg per day, about 165 μg/kg per day, about 170 μg/kg per day, about175 μg/kg per day, about 180 μg/kg per day, about 190 μg/kg per day,about 200 μg/kg per day, about 225 μg/kg per day, about 250 μg/kg perday, about 300 μg/kg per day, about 400 μg/kg per day, or about 500μg/kg per day.

A reduction in somatic cell count usually results from the effectivetreatment of inflammation and/or infection in breast or udder tissue,more particularly a mammary gland. Accordingly, a fifth aspect of thepresent invention provides a method for reducing the somatic cell countin a lactating subject comprising administering the composition or theformulation as defined above to the lactating subject. The presentinvention also provides use of a non-neutralised tocol phosphate and avitamin A compound for reducing the somatic cell count in a lactatingsubject. The present invention further provides use of a non-neutralisedtocol phosphate and a vitamin A compound in the manufacture of amedicament for reducing the somatic cell count in a lactating subject.

The lactating subject may be a female ungulate (even-toed and odd-toed),more particularly an ungulate whose milk is used for commercial purposesincluding cows, goats, sheep, pigs, yaks, water buffalo, horses,reindeer, camels, alpacas, bantengs, donkeys, oxen, zebu, moose andbison. The lactating subject may also be a lactating human.

Administration of the composition or the formulation to the lactatingsubject may reduce the somatic cell count by up to 90%, up to 80%, up to70%, up to 60%, up to 50%, up to 40%, up to 30%, up to 20%, or up to10%. The reduction may be achieved in 1 week, 2 weeks, 3 weeks, 4 weeks,5 weeks, or more, with daily or weekly administration of an effectivedose of the composition or the formulation to the lactating subject.

A composition or a formulation of the present invention may alsosupplement vitamin E levels (μmol/L) in a subject. Accordingly, thepresent invention also provides a method for supplementing vitamin Elevels in a subject comprising administering the composition or theformulation as defined above to the subject. The present invention alsoprovides use of a non-neutralised tocol phosphate and a vitamin Acompound for supplementing vitamin E is levels in a subject. The presentinvention further provides use of a non-neutralised tocol phosphate anda vitamin A compound in the manufacture of a medicament to supplementvitamin E levels in a subject.

DETAILED DESCRIPTION

The present invention relates to a composition comprising anon-neutralised tocol phosphate and a vitamin A compound.

Non-Neutralised Tocol Phosphate

The term “tocol” includes any of the naturally occurring fat-solublecompounds with vitamin E activity, namely the four tocopherols and thefour tocotrienols. The tocopherols and tocotrienols may be natural orsynthetic.

The four tocopherols and four tocotrienols occur in alpha, beta, gammaand delta forms, determined by the number and location of methyl groupson the chroman ring. The tocopherol and tocotrienol forms of Vitamin Eare shown by Formula (I):

R₁ R₂ R₃ α-tocopherol CH₃ CH₃ CH₃ α-tocotrienol CH₃ CH₃ CH₃ β-tocopherolCH₃ H CH₃ β-tocotrienol CH₃ H CH₃ γ-tocopherol H CH₃ CH₃ γ-tocotrienol HCH₃ CH₃ δ-tocopherol H H CH₃ δ-tocotrienol H H CH₃

The terms “tocopheryl phosphates” and “tocotrienyl phosphates” refer toany one of the tocopherols or tocotrienols, as shown above, where aphosphate group (PO₄) is covalently bonded via the oxygen of thehydroxyl group of tocopherol or tocotrienol.

Tocopherol or tocotrienol, in the alpha, beta, gamma or delta form, or acombination thereof, may be phosphorylated by reaction with P₄O₁₀. Thenon-neutralised tocol phosphate is the crude phosphorylation reactionproduct obtained prior to the neutralisation step.

This crude phosphorylation reaction product may comprise a number ofcomponents including both mono- and di-phosphorylated tocol phosphates.

The non-neutralised tocol phosphate may be non-neutralised tocopherylphosphates, non-neutralised tocotrienyl phosphates, or a combinationthereof.

The non-neutralised tocol phosphate may be selected from the groupconsisting of non-neutralised mono-(tocopheryl) phosphate,non-neutralised mono-(tocopheryl) phosphate monosodium salt,non-neutralised mono-(tocopheryl) phosphate disodium salt,non-neutralised mono-(tocopheryl) phosphate monopotassium salt,non-neutralised mono-(tocopheryl) phosphate dipotassium salt,non-neutralised di-(tocopheryl) phosphate, non-neutraliseddi-(tocopheryl) phosphate monosodium salt, non-neutraliseddi-(tocopheryl) phosphate monopotassium salt, non-neutralisedmono-(tocotrienyl) phosphate, non-neutralised mono-(tocotrienyl)phosphate monosodium salt, non-neutralised mono-(tocotrienyl) phosphatedisodium salt, non-neutralised mono-(tocopheryl) phosphate monopotassiumsalt, non-neutralised mono-(tocotrienyl) phosphate dipotassium salt,non-neutralised di-(tocotrienyl) phosphate, non-neutraliseddi-(tocotrienyl) phosphate monosodium salt, non-neutraliseddi-(tocotrienyl) phosphate monopotassium salt, or a combination thereof.

Other pharmaceutically or veterinary acceptable salts of the tocolphosphate may be used, such as other alkali metal salts. Otherpharmaceutically acceptable salts are well known in the art, and includethe acceptable salts described in detail in S. M. Berge, et al., J.Pharmaceutical Sciences, 66:1-19, 1977.

In some embodiments, the non-neutralised tocol phosphate is acombination of a non-neutralised neutralised mono-(tocopheryl)phosphate, non-neutralised mono-(tocopheryl) phosphate monosodium salt,non-neutralised mono-(tocopheryl) phosphate disodium salt,non-neutralised mono-(tocopheryl) phosphate monopotassium salt ornon-neutralised mono-(tocopheryl) phosphate dipotassium salt and anon-neutralised di-(tocopheryl) phosphate, non-neutraliseddi-(tocopheryl) phosphate monosodium salt or non-neutraliseddi-(tocopheryl) phosphate monopotassium salt. In other embodiments, thenon-neutralised tocol phosphate is a combination of a non-neutralisedmono-(tocotrienyl) phosphate, non-neutralised mono-(tocotrienyl)phosphate monosodium salt, non-neutralised mono-(tocotrienyl) phosphatedisodium salt, non-neutralised mono-(tocopheryl) phosphate monopotassiumsalt or non-neutralised mono-(tocotrienyl) phosphate dipotassium saltand a non-neutralised di-(tocotrienyl) phosphate, non-neutraliseddi-(tocotrienyl) phosphate monosodium salt or non-neutraliseddi-(tocotrienyl) phosphate monopotassium salt.

In one embodiment, the combination is a non-neutralisedmono-(tocopheryl) phosphate and a non-neutralised di-(tocopheryl)phosphate. It should be noted that a combination of a non-neutralisedmono-(tocopheryl) phosphate and a non-neutralised di-(tocopheryl)phosphate may be referred to herein as a non-neutralised tocopherylphosphate mixture or non-neutralised TPM.

When the composition comprises a combination, the ratio (w/w %) ofnon-neutralised mono-(tocol) phosphate to non-neutralised di-(tocol)phosphate may be at least 2:1, or about 4:1 to about 1:4, or about 6:4to about 8:2. In some embodiments, the ratio may be about 2:1, about6:4, or about 8:2.

The non-neutralised tocol phosphate has a pH of less than about 4compared to “neutralised” tocol phosphates which have a pH closer toneutral, usually in the range of above 5 to about 7. The pH of thenon-neutralised tocol phosphate may be in the range of about 2 to about4 or about 2 to about 3. Preferably, the pH of the non-neutralised tocolphosphate is about 2 or 3.

The non-neutralised tocol phosphate may comprise from about 0.01% w/w upto about 40% w/w, from about 0.01% w/w up to about 30% w/w, from about0.01% w/w up to about 20% w/w, from about 0.01% w/w up to about 10% w/w,from about 0.01% w/w up to about 5% w/w, from about 0.1% w/w up to about5% w/w, from about 0.1% w/w up to about 2.5% w/w, from about 0.1% w/w upto about 0.5% w/w, or about 0.5% w/w, about 1% w/w, about 2% w/w, about3% w/w, about 5% w/w, about 10% w/w, or about 20% w/w, of the totalconcentration of the composition.

Vitamin A Compound

The composition of the present invention also comprises a vitamin Acompound.

The vitamin A compound should be capable of converting into an activeform of vitamin A in the body and/or should be capable of providing,directly or indirectly, a compound having vitamin A activity. Thevitamin A compound may be selected from the group consisting of vitaminA, a derivative of vitamin A, a metabolite of vitamin A, a precursor ofvitamin A, or pro-vitamin A, or a combination thereof.

Vitamin A has a beta-ionone ring to which an isoprenoid chain isattached, called a retinyl group. Vitamin A is also known as retinolwhich is a member of the retinoid class.

Retinoids are considered natural or synthetic derivatives of vitamin A.The term encompasses any compound that is structurally similar toretinal (aldehyde), retinol (alcohol), or any other substance thatexhibits vitamin A activity. Retinoids may be classified as first,second and third generation retinoids. First generation retinoidsinclude retinol, retinal (retinaldehyde), tretinoin (retinoic acid),isotretinoin and alitretinoin, second generation retinoids includeetretinate and its metabolite acitrein, and third generation retinoidsinclude tazarotene, bexarotene and adapalene.

Some examples of vitamin A derivatives capable of providing, directly orindirectly, a compound having vitamin A activity, include derivatives ofretinol such as retinyl esters. Examples of retinyl esters includeretinyl acetate and retinyl palmitate.

Examples of a metabolite of vitamin A include retinoic acid and retinal.

The vitamin A compound may also be a precursor of vitamin A orpro-vitamin A.

A precursor of vitamin A is any compound that participates in thechemical reaction that produces vitamin A or a vitamin A compound thatconverts into an active form of vitamin A in the body and/or is capableof providing, directly or indirectly, a compound having vitamin Aactivity. Pro-vitamin A on the other hand is a substance that can beconverted into vitamin A by animal tissues, similar to a prodrug that ismetabolised into a drug.

Carotenoids are converted into an active form of vitamin A in the body,and thus have vitamin A activity. There are over 600 known carotenoids,which are split into two classes, xanthophylls (which contain oxygen)such as lutein and zeaxanthin and carotenes (which are purelyhydrocarbons, and contain no oxygen) such as alpha-carotene,beta-carotene, gamma-carotene, beta-cryptoxanthin and lycopene.

The amount of vitamin A compound present in the composition may beexpressed as a ratio of the amount of non-neutralised tocol phosphate tothe amount of vitamin A compound. The ratio (w/w %) of non-neutralisedtocol phosphate to vitamin A compound may be from about 0.01:1 to about100:1, from about 0.01:1 to about 5:1, from about 0.05:1 to about 2:1,from about 1:1 to about 5:1, from about 10:1 to about 60:1, or fromabout 30:1 to about 50:1. In some embodiments, the ratio may be about0.05:1, about 0.1:1, about 1:1, about 5:1, about 10:1, about 25:1, orabout 40:1.

Formulation

The present invention also relates to a formulation comprising thecomposition as defined above and a delivery vehicle.

The term “formulation” is intended to refer to the composition with adelivery vehicle and optionally other conventional excipients includingencapsulating materials such as a capsule in which the composition issurrounded by the encapsulation material. Any delivery vehicle andconventional excipients must be “pharmaceutically and/or veterinaryacceptable” meaning that it is compatible with the components of thecomposition and is not deleterious to a subject. The formulations of thepresent invention may contain other therapeutic agents, and may beformulated, for example, by employing conventional solid or liquidvehicles or diluents, as well as pharmaceutical and/or veterinaryadditives of a type appropriate to the mode of desired administration(for example, excipients, binders, preservatives, stabilisers, flavoursand so forth) according to techniques such as those well known in theart of pharmaceutical formulation (see, for example, Remington: TheScience and Practice of Pharmacy, 21st Ed., 2005 , Lippincott Williams &Wilkins).

In a preferred embodiment, the delivery vehicle is a hydrophobicdelivery vehicle.

The hydrophobic delivery vehicle may be a solid or a liquid.

Some examples of hydrophobic delivery vehicles which are most suitablefor the present invention include, but are not limited to, oils andwaxes.

Suitable oil-based hydrophobic delivery vehicles include any oil that issuitable for therapeutic use, such as for example, any edible oil. Theoil-based hydrophobic delivery vehicles may be natural or synthetic. Theoil-based hydrophobic delivery vehicles should also be compatible withthe non-neutralised tocol phosphate and the vitamin A compound.

Preferred oil-based hydrophobic delivery vehicles suitable for use inthe composition of the present invention include vegetable oil, fruitoil, seed oil, grain oil, nut oil, or the like. These oils may besaturated or unsaturated. These oils may also be winterised ornon-winterised. Examples of suitable oils include canola oil, coconutoil, corn oil, cottonseed oil, olive oil, Olea europaea (olive) leafextract, palm oil, peanut oil, rapeseed oil, safflower oil, sesame oil,soybean oil, sunflower oil, almond oil, cashew oil, hazelnut oil,macadamia oil, Macadamia integrifolia (macadamia) seed oil, mongongo nutoil, pecan oil, pine nut oil, pistachio oil, walnut oil, bottle gourdoil, buffalo gourd oil, pumpkin seed oil, watermelon seed oil, acai oil,blackcurrant seed oil, borage seed oil, evening primrose oil (Oenotherabiennis oil), amaranth oil, apricot oil, apple seed oil, argan oil,artichoke oil, avocado oil (Persea gratissima oil), babassu oil, benoil, borneo tallow nut oil, cape chestnut oil, cocoa butter, carob podoil, cocklebur oil, cohune oil, coriander seed oil, dika oil, false flaxoil, flax seed oil, grape seed oil (Vitis vinifera), hemp oil, kapokseed oil, lallemantia oil, marula oil, meadowfoam seed oil, mustard oil,nutmeg butter, okra seed oil, orange oil, papaya seed oil, perilla seedoil, pequi oil, pine nut oil, poppyseed oil, prune kernel oil, quinoaoil, ramtil oil, rice bran oil (for example, Oryza Satiraυ Bran Oil),rose hip oil (Rosa eglanteria oil), royle oil, sacha inchi oil,sandalwood oil (Santalum spicatum oil), tea seed oil, thistle oil,tomato seed oil and wheat germ oil.

Suitable hydrophobic delivery vehicles may also include cetearylalcohol, cetearyl glucoside, cetearyl olivate, cocoyl proline, dicaprylether, glycerin, glyceryl linoleate, glyceryl oleate, lactic acid,lecithin, pomegranite sterols, resveratrol and vitamin D3.

Suitable hydrophobic delivery vehicles may also include polyunsaturatedoils containing polyunsaturated fatty acids. In one or more embodiments,the unsaturated fatty acids are selected from the group of omega-3 andomega-6 fatty acids. Examples of such polyunsaturated fatty acids arelinoleic and linolenic acid, gamma-linoleic acid (GLA), eicosapentaenoicacid (EPA) and docosahexaenoic acid (DHA). In some instances, mixturesof fatty acids or medium chain triglycerides are excluded from the scopeof hydrophobic delivery vehicles.

Another class of hydrophobic delivery vehicles is essential oils, whichmay also be considered therapeutically-active oils if they containactive biologically occurring molecules and exert a therapeutic effecton administration. In the context of the present invention, deliveryvehicles that additionally possess therapeutically beneficial propertiesmay be referred to as “therapeutically-active delivery vehicle”. If aformulation is made using a therapeutically-active delivery vehicle,then the formulation will include a non-neutralised tocol phosphate anda vitamin A compound as well as the therapeutically-active deliveryvehicle. Non-limiting examples of therapeutically-active deliveryvehicles include essential oils such as rosehip oil and tea tree oil.Other examples of essential oils are oils of anise, basil, bergamot,camphor, cardamom, carrot, canola, cassia, catnip, cedarwood,citronella, clove, cypress, eucalyptus, frankincense, garlic, ginger,grapefruit, hyssop, jasmine, jojoba, lavender, lavandin, lemon, lime,mandarin, marjoram, myrrh, neroli, nutmeg, orange, peppermint,petitgrain, rosemary, sage, spearmint, star anise, tangerine, thymevanilla, verbena and white clover.

Another class of therapeutically-active oils includes liquid hydrophobicplant derived oils, which are known to possess therapeutic benefits whenapplied topically.

Silicone oils also may be used and may be particularly desirable due totheir known skin protective and occlusive properties. Suitable siliconeoils include non-volatile silicones, such as polyalkyl siloxanes,polyaryl siloxanes, polyalkylaryl siloxanes and polyether siloxanecopolymers, polydimethylsiloxanes (dimethicones) andpoly(dimethylsiloxane)(diphenyl-siloxane) copolymers. Silicone oils mayalso considered therapeutically-active oil due to their barrierretaining and protective properties.

Hydrophobic liquids selected from the family of organic liquidsdescribed as “emollients” is another class of hydrophobic deliveryvehicles. Emollients possess a softening or soothing effect, especiallywhen applied to body areas, such as the skin and mucosal surfaces.Examples of suitable emollients include isopropyl myristate, isopropylpalmitate, isopropyl isostearate, diisopropyl adipate, diisopropyldimerate, maleated soybean oil, octyl palmitate, cetyl lactate, cetylricinoleate, tocopheryl acetate, cetyl acetate, tocopheryl linoleate,wheat germ glycerides, arachidyl propionate, myristyl lactate, decyloleate, propylene glycol ricinoleate, isopropyl lanolate,pentaerythrityl tetrastearate, neopentylglycol dicaprylate/dicaprate,isononyl isononanoate, isotridecyl isononanoate, myristyl myristate,octyl dodecanol, sucrose esters of fatty acids and octylhydroxystearate.

Suitable hydrophobic delivery vehicles also include pharmaceuticallyacceptable waxes. An example of a pharmaceutically acceptable wax isbeeswax.

It should be noted that, as used in the present specification, thesingular forms “a”, “an” and “the” include plural aspects unless thecontext clearly dictates otherwise. Thus, for example, reference to “ahydrophobic delivery vehicle” may refer to a single hydrophobic deliveryvehicle, or two or more hydrophobic delivery vehicles, depending on thecontext, and so forth for other features of the invention.

The delivery vehicle may be present in an amount of at least about 60.0%w/w, at least about 80.0% w/w, at least about 90% w/w, at least about99.0% w/w, or at least about 99.5% w/w, of the total concentration ofthe formulation. In some embodiments, the delivery vehicle is present inan amount of about 60.0% w/w, about 80.0% w/w, about 90% w/w, about 98%w/w, about 99% w/w, or about 99.5% w/w, of the total concentration ofthe formulation.

Nutrient Compound

The composition or the formulation may optionally comprise a nutrientcompound.

The term “nutrient compound” encompasses antioxidants, vitamins,minerals and trace elements which are capable of treating inflammationand/or infection in breast or udder tissue of a subject, reducing thesomatic cell count in a lactating subject or supplementing vitamin Elevels in a subject.

The nutrient compound may be selected from the group consisting of, butis not limited to, coenzyme Q10 (or ubiquinone); ubiquinol; fat-solublevitamins such as vitamin D compounds (e.g. D2, D3 and theirderivatives), vitamin E compounds (α, β, γ, δ-tocopherols, or α, β, γ,δ-tocotrienols), vitamin K compounds (e.g. K1, K2, K3 and theirderivatives); folic acid; vitamin B compounds (e.g. B1, B2, B3, B6 andB12); vitamin C; vitamin P; vitamin F; lutein; zeaxanthin; cysteine;flavonoids (e.g. catechins, reserveratrol, proanthocyanidins);isoflavones (e.g. genistein and daidzein); bilberry; ginkgo biloba;grape seed extract; phytonutrients (e.g. lycopene, lutein andseaxanthin); alpha lipoic acid; bilberry; bioflavinoids; unsaturatedfatty acids (e.g. linoleic acid, conjugated linoleic acid, linolenicacid, omega-3 fatty acids such as docosahexaenoic acid (DHA) andeicosapentaeonic acid (EPA) and their glycerol-esters); calcium;phosphorus; magnesium; fluorine; phosphorus; sulfur; sodium; potassium;chloride; calcium; iodine; cobalt; copper; iron; manganese; molybdenum;selenium; zinc; chromium; cadmium; fluorine; nickel; silicon; tin;vanadium; niacin; and combinations thereof.

Excipients

A formulation of the present invention can optionally further compriseone or more excipients. A person skilled in the art of the inventionwould appreciate suitable excipients that could be included informulations of the present invention. The choice and amount ofexcipients will depend on the intended use of formulations, the mode ofadministration and/or the dosage form.

Examples of suitable excipients include additional solvents such aswater, thickeners or gelling agents, surfactants, buffers, emollients,sweeteners, disintegrators, flavours, colours, fragrances, electrolytes,appearance modifiers, film foaming polymers and the like. Suitablesweeteners include sucrose, lactose, glucose, aspartame or saccharin.Suitable disintegrators include corn starch, methylcellulose,polyvinylpyrrolidone, xanthan gum, bentonite, alginic acid or agar.Suitable flavours include peppermint oil, oil of wintergreen, cherry,orange, or raspberry flavouring. Other flavourings may be molasses,salt-licks, and the like, which are particularly suitable for animals.Suitable preservatives include sodium benzoate, methylparaben,propylparaben, and sodium bisulphite. However, it will be appreciatedthat any excipients, which have been approved for use in pharmaceuticaland veterinary products by the regulatory bodies, may be employed in theformulations of the present invention.

Preparation of Compositions and Formulations

The compositions and the formulations of the present invention may beprepared by a variety of techniques.

One method of preparing the composition defined above comprises thesteps of mixing a non-neutralised tocol phosphate and a vitamin Acompound, in suitable quantities, with stirring, until completehomogenisation is achieved. Another method of preparing the compositiondefined above comprises the steps of warming the non-neutralised tocolphosphate to a temperature greater than about 60° C., preferably betweenabout 70° C. and about 80° C., and then adding the a vitamin A compoundto the non-neutralised tocol phosphate when the non-neutralised tocolphosphate is at a temperature of less than about 40° C., with stirring,until complete homogenisation is achieved. One method for preparing theformulation as defined above comprises the steps of mixing thenon-neutralised tocol phosphate and the hydrophobic delivery vehicle,and warming the mixture to a temperature greater than about 60° C.,preferably between about 70° C. and about 80° C., and then adding thevitamin A compound to the non-neutralised tocol phosphate when thenon-neutralised tocol phosphate is at a temperature of less than about40° C., with stirring, until complete homogenisation is achieved.

Treatment of Inflammation and/or Infection in Breast or Udder Tissue

The compositions and the formulations of the present invention aresuitable for the treatment of inflammation and/or infection in breast orudder tissue, more particularly in a mammary gland. The inflammationand/or infection in a mammary gland may be mastitis.

The “subject” may be mammal including humans and animals. In oneembodiment, the mammal is a human, female or male. In other embodiments,the mammal is an animal, in particular of the subclass Theria includingthe subclasses Metatheria and Eutheria. The mammal may also be amonotreme of the subclass Prototheria. However, it is recognised thatthese are mammals in which the mammary glands are devoid of teats. Thisis also the case for a monotremata of the subclass Australospenida whichincludes the platypus and the echidna.

Under the subclass of Metatheria are the superorders of Ameridelphia andAustralidelphia which are inclusive of marsupials such as kangaroos andpossums. The subclass of Eutheria is inclusive of placentals. Under thesubclass of Eutheria are a number of superorders, which in turnencompass orders. The superorder of Xenarthra includes the ordersCingulata (armadillos) and Pilosa (anteaters and sloths). The superorderof Afrotheria includes the orders Afrosoricida (tenrecs and goldenmoles), Macroscelidea (elephant shrews), Tubulidentata (aardvarks),Hyracoidea (hyraxes), Proboscidea (elephants) and Sirenia (dugongs andmanatees). The superorder of Laurasiatheria includes the ordersSoricomorpha (shrews, moles and solenodons), Erinaceomorpha (hedgehogsand relatives), Chiroptera (bats), Pholidota (pangolins), Carnivora(dogs, cats, weasels, bears, seals, and their relatives), Perissodactyla(odd-toed ungulates), Artiodactyla (even-toed ungulates) and Cetacea(whales and dolphins), the latter two orders are sometimes referred tothe order of Cetartiodactyla. The superorder of Euarchontogliresincludes the orders Rodentia (mice, rats, porcupines, beavers,capybaras, and other gnawing mammals), Lagomorpha (rabbits andrelatives), Scandentia (treeshrews), Dermoptera (colugos) and Primates(humans, apes and monkeys).

In one embodiment, the subject may be selected from humans, domesticmammals such as companion animals, working animals, livestock, andzoological/park mammals.

Preferably, the subject is a mammal, in particular a female mammal, moreparticularly a female mammal having developed lactiferous ducts such asa lactating female mammal.

These mammals include female humans, whose milk is needed for feedingtheir young, or expressed and donated to milk banks for redistributionto infants who may benefit from human milk for various reasons (e.g.premature neonates, babies with allergies, and metabolic diseases). Themammals also include female ungulates (even-toed and odd-toed) whosemilk is used for commercial purposes, particularly as a food product forhuman consumption (e.g. cows, goats, sheep, yaks, water buffalos,horses, reindeer, camels, alpacas, bantengs, donkeys, oxen, zebu, mooseand bison). The mammals may also include other lactating mammals whichcould benefit from the present invention such as those under veterinarycare (e.g. domestic mammals including livestock, companion animals andworking animals, or zoological/park mammals such as apes, monkeys,lions, zebras, and so on).

There are a number of causes for inflammation and/or infection in breastor udder tissue, more particularly in a mammary gland, which can occurin the subjects defined above. It may be caused by physical damage tobreast or udder tissue such as by injury produced by physical force,chemicals introduced into the mammary gland or from bacteria and theirtoxins entering the breast or udder tissue.

Inflammation and/or infection in breast or udder tissue, moreparticularly in a mammary gland, are often associated with mastitis.

There are two types of mastitis: puerperal mastitis and non-puerperalmastitis. It is called puerperal mastitis when it occurs in lactatingmothers, and non-puerperal otherwise. Mastitis can also occur in men,albeit rarely. Puerperal mastitis is the inflammation of the breast inconnection with pregnancy, breastfeeding or weaning. Puerperal mastitisis thought to be caused by blocked milk ducts or milk excess. Theincidence of puerperal mastitis is estimated to be in the range of 5-33%of breastfeeding mothers. The term non-puerperal mastitis describesinflammatory lesions of the breast occurring unrelated to pregnancy andbreastfeeding. Non-puerperal mastitis may also be referred to as plasmacell mastitis, subareolar abscess, duct ectasia, periductalinflammation, Zuska's disease, and others.

Infections play only a minor role in the pathogenesis of human mastitis,with many cases of mastitis being aseptic under normal hygienicconditions. However, infection is a primary cause of mastitis inveterinary mastitis and in poorly hygienic conditions.

Where bacterial infection is involved, they are usually caused by acommon bacterium found on the skin, inside the udder of animals, or onthe skin of the teat. These bacteria are typically Staphylococcus aureusand Streptococcus agalactiae, but there are numerous others. Thebacteria will usually enter breast or udder tissue through a break orcrack in the skin. The infection then takes place in the tissuesurrounding the milk ducts, causing swelling which compresses on themilk ducts, and pain in the infected breast.

Accordingly, the present invention provides a method for the treatmentof inflammation and/or infection in breast or udder tissue comprisingadministering the composition or the formulation as defined above to asubject in need thereof. The present invention also provides use of anon-neutralised tocol phosphate and a vitamin A compound for thetreatment of inflammation and/or infection in breast or udder tissue.The present invention further provides use of a non-neutralised tocolphosphate and a vitamin A compound in the manufacture of a medicamentfor the treatment of inflammation and/or infection in breast or uddertissue. The breast or udder tissue may be a mammary gland.

Generally, the term “treating” means affecting a subject, tissue or cellto obtain a desired pharmacological and/or physiological effect andincludes: (a) inhibiting the inflammation and/or infection in breast orudder tissue, such as by arresting its development or furtherdevelopment; (b) relieving or ameliorating the effects of theinflammation and/or infection in breast or udder tissue, such as bycausing regression of the effects of the inflammation and/or infectionin breast or udder tissue; (c) reducing the incidence of theinflammation and/or infection in breast or udder tissue, or (d)preventing the inflammation and/or infection in breast or udder tissuefrom occurring in a subject, tissue or cell predisposed to theinflammation and/or infection in breast or udder tissue, or at riskthereof, but has not yet been diagnosed with a protectivepharmacological and/or physiological effect so that the inflammationand/or infection in breast or udder tissue does not develop or occur inthe subject, tissue or cell.

Although the present invention has been described with reference totreating inflammation and/or infection in breast or udder tissue, itwill be appreciated that the present invention may also be useful in thetreatment of other diseases or disorders associated with theinflammation and/or infection in breast or udder tissue such as, forexample, inflammatory breast cancer, which has symptoms very similar tomastitis.

As mastitis occurs in domestic mammals as in humans, it is especially aconcern in livestock, since milk from the affected udders of livestockmay enter the food supply and pose a health risk. It is a majorcondition in some species, like dairy cows, because of the tremendouseconomic importance for the dairy industry. The same considerationsapply to mastitis in sheep and goats and other milk producing livestock.It is also of economic importance in the sow, but, in this species, itis not related to public health. In other mammals, it is more anindividual illness dealt with by veterinary practitioners.

There are several ways of classifying mastitis in domestic mammals. Asimple classification recognises mastitis in two major groups: (a)contagious mastitis, which is caused by bacteria live on the skin of theteat and inside the udder and can be transmitted, for example, from onedairy cow to another during milking, and (b) environmental mastitis,which describes mastitis caused by organisms such as Escherichia coliwhich do not normally live on the skin or in the udder but which enterthe teat canal when in contact with a contaminated environment. Thepercentage of cases of environmental mastitis is quite small compared tothe total mastitis cases in domestic mammals.

Contagious mastitis can be divided into three groups: (i) clinicalmastitis, (ii) sub-clinical mastitis, and (iii) chronic mastitis.Clinical mastitis is characterised by the presence of gross inflammationsigns (swelling, heat, redness, pain). Three types of clinical mastitisexist. Peracute mastitis characterised by gross inflammation, disruptedfunctions (reduction in milk yield, changes in milk composition) andsystemic signs (fever, depression, shivering, loss of appetite and lossof weight). Acute mastitis is similar to peracute mastitis, but withlesser systemic signs (fever and mild depression). Sub-acute mastitiswhen the mammary gland inflammation signs are minimal and no visiblesystemic signs. Sub-clinical mastitis is characterised by change in milkcomposition with no signs of gross inflammation or milk abnormalities.Changes in milk composition can be detected by special diagnostic tests.Chronic mastitis is where the inflammatory process exists for months,and may continue from one lactation period to another. Chronic mastitisfor the most part exists as sub-clinical but may exhibit periodicflare-ups sub-acutely or acutely, which last for a short period of time.

Clinical and sub-clinical mastitis are the most important forms and canbe diagnosed on the basis of bacteriological examination or by indirecttests, principally based on the somatic cell count, which is one of theindicators of the quality of milk. It should be noted that humans mayalso experience clinical and sub-clinical mastitis, however the milk ofthese subjects is usually not subjected to somatic cell count fordiagnosis.

Somatic cells are leukocytes (white blood cells) and can containlipolytic and proteolytic enzymes, which degrade fats and proteins,respectively. An increase in somatic cells count during a mastitisinfection increases the amount of destructive enzymes present in themilk, which increases the rate of deterioration of the milk fat andprotein. The number of somatic cells increases in response to pathogenicbacteria like Staphylococcus aureus, a cause of mastitis. The somaticcell count is usually quantified as cells per ml. With respect to dairycows, it is generally considered that a somatic cell count less than100,000 cells/ml is “uninfected”, whereas a somatic cell count ofgreater than 100,000 cells/ml and up to 500,000 cells/ml is likely to besub-clinical mastitis and a higher somatic cell count, that is greaterthan 500,000 cells/ml, is likely to be clinical mastitis.

The compositions and the formulations of the present invention have beenshown to reduce the somatic cell count in livestock and therefore thecompositions and the formulations of the present invention provide aneffective the treatment of mastitis, particularly clinical andsub-clinical mastitis.

Accordingly, the present invention also provides a method for reducingthe somatic cell count in a lactating subject comprising administeringthe composition or the formulation as defined above to the lactatingsubject. The present invention also provides use of a non-neutralisedtocol phosphate and a vitamin A compound for reducing the somatic cellcount of a lactating subject. The present invention further provides useof a non-neutralised tocol phosphate and a vitamin A compound in themanufacture of a medicament for reducing the somatic cell count in alactating subject.

In a preferred embodiment, the lactating subject is a female ungulate(even-toed and odd-toed), more particularly a female ungulate whose milkis used for commercial purposes including cows, goats, sheep, pigs,yaks, water buffalo, horses, reindeer, camels, alpacas, bantengs,donkeys, oxen, zebu, moose and bison. The lactating subject may also bea lactating human.

Administration of the composition or the formulation to the lactatingsubject may reduce the somatic cell count by up to 90%, up to 80%, up to70%, up to 60%, up to 50%, up to 40%, up to 30%, up to 20%, or up to10%. The reduction may be achieved in 1 week, 2 weeks, 3 weeks, 4 weeks,5 weeks, or more, with daily or weekly administration of an effectivedose of the composition or the formulation to the lactating subject.

Vitamin E Supplementation

A composition or a formulation of the present invention may alsosupplement vitamin E levels (μmol/L) in a subject. Accordingly, thepresent invention also provides a method for supplementing vitamin Elevels in a subject comprising administering the composition or theformulation as defined above to the subject. The present invention alsoprovides use of a non-neutralised tocol phosphate and a vitamin Acompound for supplementing vitamin E levels in a subject. The presentinvention further provides use of a non-neutralised tocol phosphate anda vitamin A compound in the manufacture of a medicament to supplementvitamin E levels in a subject.

Administration Route

Routes of administration can broadly be divided into a three categoriesby effect, namely, “topical” where the desired effect is local, so thesubstance is applied directly where its action is desired, “enteral”where the desired effect is systemic (non-local) so the substance isgiven via the digestive tract, and “parenteral” where the desired effectis systemic, so the substance is given by routes other than thedigestive tract.

The formulation of the present invention is suitable for topical,enteral or parenteral administration.

Examples of topical routes of administration having a local effectinclude epicutaneous (onto the skin) including the skin of the breast ofa human subject or the udder of an animal subject.

Examples of enteral routes of administration having a systemic(non-local) effect include any form of administration that involves anypart of the gastrointestinal tract, such as oral (into the mouth),intranasal (into the nose), rectal (into the rectum), and vaginal (intothe vagina). Oral administration includes buccal administration(absorbed through the cheek near the gumline), and sublingualadministration (under the tongue).

Examples of parenteral routes of administration by injection, infusionor diffusion having a systemic effect include intravenous (into a vein),intraarterial (into an artery), intramuscular (into a muscle),intracardiac (into the heart), subcutaneous (under the skin),percutaneous (via needle-puncture into the skin), intradermal (into theskin itself), intrathecal (into the spinal canal), intraperitoneal(infusion or injection into the peritoneum), intravesical infusion(infusion into the urinary bladder), epidural (injection or infusioninto the epidural space), transdermal or transcutaneous (diffusionthrough the intact skin), transmucosal (diffusion through a mucousmembrane), insufflation (diffusion through the nose), inhalational(diffusion through the mouth), and intramammary (into mammary tissue).

Topical, in particular epicutaneus, enteral, in particular oral, andparental, in particular intramammary, routes of administration arepreferred.

Dosage Form

The compositions and the formulations of the present invention may beprepared into any suitable dosage form for topical, enteral, parenteraladministration.

A person skilled in the art would readily appreciate what would be asuitable dosage form for topical, enteral, parenteral administration

Suitable dosage forms for topical administration include creams,lotions, gels and the like. The dosage form may also be a patch or otherdevice. As one example for veterinary applications, the device could bein the form of a cover for the teat such as a Teatseal®, which is placedonto the teat after milking as a barrier to bacterial contamination. Inthis application, the composition or the formulation of the presentinvention could be provided on the inner surface of such a teat cover sothat the composition or the formulation contacts the teat when in place.

Suitable dosage forms for enteral administration would include but notbe limited to capsules, tablets, pills, or specialty tablets such asbuccal, sublingual, chewable tablets or orally-disintegrating tablets.Another example of a suitable dosage form would be edible thin films.

Other suitable dosage forms for enteral administration include liquidsolutions or suspensions. Suitable liquid solution or suspension dosageforms may be in the form of a drink, such as sports drinks containingelectrolytes (e.g. gatorade), or syrup and elixirs. Other suitableliquid solution or suspension dosage forms include nasal deliverysolutions and oral suspensions including liquid solutions orsuspensions. For veterinary purposes, the liquid solution or suspensionmay be in the form of a “drench”. The liquid solution or suspension mayalso be used in the preparation of an edible product, such as a biscuitor cake, suitable for human or animal consumption. A product for animalconsumption may also include appropriate animal feeds.

The dosage form for enteral administration may also be a powder or solidcrystal, which can be either dissolved or suspended in a liquid beforeadministration. Alternatively, the powder may be consumed directly oradded to a food or drink product for consumption. In the case of farmanimals, the formulation may be added directly to the animal feed.

Where the composition or the formulation has a disagreeable taste,additives with sufficient flavour to disguise the bad taste may be addedto the dosage form (e.g. masking agents).

Examples of suitable dosage forms for parenteral administration includebut are not limited to injectables (i.e. solutions, suspensions,emulsions, and dry powders for reconstitution), intramammary infusions,intravaginal delivery systems, and implants.

Dosage Regime

The term “therapeutically effective amount” refers to the amount of acomposition or a formulation of the present invention that will elicitthe biological or medical response of a subject, tissue or cell that isbeing sought by the researcher, veterinarian, medical doctor or otherclinician. Administration of a “therapeutically effective amount” of thecomposition or the formulation of the present invention should obtain adesired pharmacological and/or physiological effect and includes: (a)inhibiting the inflammation and/or infection in breast or udder tissue,such as by arresting its development or further development; (b)relieving or ameliorating the effects of the inflammation and/orinfection in breast or udder tissue, such as by causing regression ofthe effects of the inflammation and/or infection in breast or uddertissue; (c) reducing the incidence of the inflammation and/or infectionin breast or udder tissue, or (d) preventing the inflammation and/orinfection in breast or udder tissue from occurring in a subject, tissueor cell predisposed to the inflammation and/or infection in breast orudder tissue, or at risk thereof, but has not yet been diagnosed with aprotective pharmacological and/or physiological effect so that theinflammation and/or infection in breast or udder tissue does not developor occur.

It will be understood that the specific dose level and frequency ofdosage for any particular subject may be varied and will depend upon avariety of factors including the activity of the composition orformulation employed, and the metabolic stability and length of actionof that composition or the formulation; the age, body weight, generalhealth, sex, or diet of the subject; the route of administration used;the mode and time of administration of the composition or theformulation; the rate of excretion or clearance of the composition orthe formulation from the body; the non-neutralised tocol phosphate andthe vitamin A compound used in the composition or the formulation; theseverity of the inflammation or infection of the breast or udder tissueof the subject; and the particular subject undergoing treatment.

Suitable intervals of dosing include monthly, every two months orlonger, biweekly, weekly, daily, or multiple times per day. Preferably,the composition or the formulation of the present invention isadministered weekly, daily, or multiple times per day.

The composition or the formulation will generally contain an appropriatedosage level of non-neutralised tocol phosphate, which may be about 0.1to about 20 mg per kg subject body weight per day which can beadministered in single or multiple doses. Preferably, the dosage levelwill be about 0.1 to about 15 mg/kg per day; about 0.1 to about 10 mg/kgper day; more preferably, about 0.1 to about 10 mg/kg per day, about 0.1to about 5 mg/kg per day, about 0.1 to about 2.5 mg/kg per day or about0.1 to about 2 mg/kg per day. A suitable dosage level may be about 0.1to about 7.5 mg/kg per day. For example, within the above dosage ranges,the dosage may be about 0.1 mg/kg per day, about 0.2 mg/kg per day,about 0.4 mg/kg per day, about 0.6 mg/kg per day, about 0.7 mg/kg perday, about 0.8 mg/kg per day, about 0.9 mg/kg per day, about 1 mg/kg perday, about 1.2 mg/kg per day, about 1.4 mg/kg per day, about 1.5 mg/kgper day, about 1.6 mg/kg per day, about 1.7 mg/kg per day, about 1.8mg/kg per day, about 1.9 mg/kg per day, about 2 mg/kg per day, about 2.2mg/kg per day, about 2.5 mg/kg per day, about 3 mg/kg per day, about 5mg/kg per day, about 7.5 mg/kg per day, or about 10 mg/kg per day.

The composition or the formulation will generally contain an appropriatedosage level of the vitamin A compound, which may be about 10 to about1000 μg per kg subject body weight per day which can be administered insingle or multiple doses. Preferably, the dosage level will be about 10to about 850 μg/kg per day; about 10 to about 800 μg/kg per day; about10 to about 500 μg/kg per day; or less than about 300 μg/kg per day.More preferably, the dosage level will be about 10 to about 500 μg/kgper day, 10 to about 275 μg/kg per day, 10 to about 250 μg/kg per day,20 to about 250 μg/kg per day, 25 to about 250 μg/kg per day, 10 toabout 200 μg/kg per day or 50 to about 200 μg/kg per day. A suitabledosage level may be about 10 to about 300 μg/kg per day. For example,within the above dosage ranges, the dosage may be about 10 μg/kg perday, about 20 μg/kg per day, about 40 μg/kg per day, about 50 μg/kg perday, about 60 μg/kg per day, about 70 μg/kg per day, about 75 μg/kg perday, about 80 μg/kg per day, about 85 μg/kg per day, about 90 μg/kg perday, about 95 μg/kg per day, about 100 μg/kg per day, about 125 μg/kgper day, about 150 μg/kg per day, about 155 μg/kg per day, about 160μg/kg per day, about 165 μg/kg per day, about 170 μg/kg per day, about175 μg/kg per day, about 180 μg/kg per day, about 190 μg/kg per day,about 200 μg/kg per day, about 225 μg/kg per day, about 250 μg/kg perday, about 300 μg/kg per day, about 400 μg/kg per day, or about 500μg/kg per day.

In one embodiment, the composition or the formulation, will beadministered daily and will comprise a tocol phosphate in an amount ofabout 0.1 to about 15 mg/kg, and a vitamin A compound in an amount ofabout 10 to about 500 μg/kg. In cases of sub-clinical mastitis, thesubject may be administered a daily dose of the composition or theformulation comprising a non-neutralised tocol phosphate in an amount ofabout 0.1 to about 1.5 mg/kg and a vitamin A compound in an amount ofabout 10 to about 150 μg/kg. In the cases of clinical mastitis, thesubject may be administered a daily dose of the composition or theformulation with increased amounts of a non-neutralised tocol phosphateand a vitamin A compound. For example, the composition or theformulation may comprise a non-neutralised tocol phosphate in an amountof about 1.0 to about 5.0 mg/kg and a vitamin A compound in an amount ofabout 100 to about 300 μg/kg.

In one embodiment, an effective dosage form comprises non-neutralisedTPM and a vitamin A compound such as beta-carotene in any of the abovedefined amounts.

FIGURES

The invention is described further by way of example with reference tothe accompanying drawings in which:

FIG. 1A is a bar chart showing the average somatic cell count of thedairy cows of Group A over a period of time prior to treatment withFormulation A (represented in grey) and over a 2 week treatment period(represented in black).

FIG. 1B is a bar chart showing the percentage change in the averagesomatic cell count of the dairy cows of Group A over a period prior totreatment with Formulation A and over a 2 week treatment period.

FIG. 2A is a bar chart showing the average somatic cell count of thedairy cows of Group B over a period of time prior to treatment withFormulation B (represented in grey) and over a 5 week treatment period(represented in black).

FIG. 2B is a bar chart showing the percentage change in the averagesomatic cell count of the dairy cows of Group B over a period of timeprior to treatment with Formulation B and over a 5 week treatmentperiod.

EXAMPLES

The invention is described further by reference to the followingnon-limiting examples of the invention.

Example 1 Preparation of Formulations

The following formulations of the present invention were prepared fororal delivery.

Component Formulation A Formulation B Non-neutralised TPM 1% w/w (~0.5g) 2% w/w (~1 g) Beta-carotene 0.1% w/w (~50 mg)   0.2% w/w (~100 mg)Olive oil balance to 100% balance to 100% (total ~60 ml) (total ~60 ml)

Each of Formulations A and B were prepared by combining non-neutralisedTPM (pH about 2 to about 4) with olive oil, and stirring the mixture atabout 70° C. The mixture was then cooled to below 40° C. before thebeta-carotene was added, with stirring.

Example 2 Treatment of Dairy Cows Suffering Mastitis

35 dairy cows showing signs of either sub-clinical or clinical mastitiswith persistent elevated somatic cell counts, were kept separated fromthe main herd (consisting of approximately 300 head). Various attemptsto treat the mastitis in the cattle had been made, and the mastitis wasconsidered problematic.

18 of the 35 dairy cows were selected based on their somatic cellcounts; their somatic cell counts and previous antibiotic treatment wasdocumented. The 18 dairy cows were divided into 2 groups consisting of 9dairy cows each and designated Group A and Group B.

Final antibiotic treatment ceased at least 4 weeks prior to commencingtreatment with formulations of the present invention. A baseline ortreatment week 0 somatic cell count was measured.

At treatment week 0, the Group A dairy cows had elevated somatic cellcounts but not severe, with an average somatic cell count of 429,000cells/ml, whereas the Group B dairy cows showed some clinical signs ofmastitis and generally had much higher somatic cell counts with anaverage somatic cell count of 1,243,222 cells/ml.

The Group A dairy cows were treated with Formulation A of Example 1 andthe Group B dairy cows were treated with Formulation B of Example 1.

Both groups were treated daily using an oral drench, administered eachmorning after milking. The oral drench was delivered via a drenching gunthat delivered approximately 60 ml of the relevant formulation.

Each week, a milk sample was collected from each of the dairy cows inthe respective groups and somatic cell counts were measured for eachindividual dairy cow.

Group A Results

The Group A dairy cows were treated for 2 weeks. The results of thisstudy are shown in the table below.

TABLE 1 Somatic cell counts for the Group A dairy cows Average Averagepercentage somatic cell change in count somatic cell Time point(cells/ml) count Previous 6 (reading taken 33 weeks before 311,000Treatment Week 0) Previous 5 (reading taken 28 weeks before 128,400Treatment Week 0) Previous 4 (reading taken 23 weeks before 222,000Treatment Week 0) Previous 3 (reading taken 17 weeks before 161,000Treatment Week 0) Previous 2 (reading taken 12 weeks before 322,444Treatment Week 0) Previous 1 (reading taken 7 weeks before 271,000Treatment Week 0) Treatment Week 0 429,000 Treatment Week 1 341556 −23%Treatment Week 2 222,111 −45%

The results shown for the somatic cell counts are an average of thesomatic cell counts of the 9 dairy cows for this group. The percentagechange values were calculated as the difference in the somatic cellcount compared to the somatic cell count at treatment week 0, andexpressed as a percentage of the somatic cell count for that treatmentweek.

The graph shown in FIG. 1A shows the decrease in somatic cell count overthe treatment period. After 2 weeks of daily treatment, the averagesomatic cell count decreased from 429,000 cells/ml to 222,111 cells/ml,which correlates to a 45% reduction in the somatic cell count after 2weeks of treatment.

Group B Results

The Group B dairy cows were treated for 5 weeks. The results of thisstudy are shown in the table below.

TABLE 2 Somatic cell counts for the Group B dairy cows Average Averagepercentage somatic cell change in count somatic cell Time point(cells/ml) count Previous 6 (reading taken 33 weeks before 1,491,000Treatment Week 0) Previous 5 (reading taken 28 weeks before 604,857Treatment Week 0) Previous 4 (reading taken 23 weeks before 221,375Treatment Week 0) Previous 3 (reading taken 17 weeks before 234,000Treatment Week 0) Previous 2 (reading taken 12 weeks before 560,111Treatment Week 0) Previous 1 (reading taken 7 weeks before 1,122,667Treatment Week 0) Treatment Week 0 1,243,222 Treatment Week 1 2,247,333109% Treatment Week 2 1,145,444 12% Treatment Week 3 1,265,556 7%Treatment Week 4 852,625 −33% Treatment Week 5 516,556 −54%

The results shown for the somatic cell counts are an average of thesomatic cell counts of the 9 dairy cows for this group. The percentagechange values were calculated as the difference in the somatic cellcount compared to the somatic cell count at treatment week 0, andexpressed as a percentage of the somatic cell count for that treatmentweek.

The graph shown in FIG. 2A shows the decrease in somatic cell count overthe treatment period. After 5 weeks of daily treatment, the averagesomatic cell count decreased from 1,243,222 cells/ml to 516,556cells/ml, which correlates to a 54% reduction in the somatic cell countafter 5 weeks of treatment.

This example clearly demonstrates the effectiveness of formulations ofthe present io invention in the reduction of somatic cell count andtherefore treatment of mastitis in dairy cows.

Example 3 Vitamin E Supplementation in Dairy Cows Suffering Mastitis

The following are results of vitamin E levels in 4 dairy cows showingsigns of mastitis treated with a formulation of the present inventionafter 1 week. The blood of the 4 dairy cows was tested at a commercialpathology laboratory.

Baseline Vitamin E Supplementation (μmol/L) Vitamin E (μmol/L) Change %Change DC1 13.6 13.9 +0.3 +2.2 DC2 9.6 10.8 +1.2 +12.5 DC3 14.6 15.2+0.6 +4.1 DC4 16.1 16.7 +0.6 +3.7

Example 4 Formulations Comprising Nutrients

Further embodiments of an oral drench can be prepared according toExample 1, but comprising additional nutrients, such as for example,selenium, copper, zinc, a vitamin E compound. The nutrients are added atthe same time as the beta-carotene.

Specifically designed oral drenches would be administered to dairycattle as described in Example 2, and compared with dairy cattleadministered with an oral drench not comprising any nutrients. The bloodlevels of vitamin E could also be measured as in Example 3.

Example 5 Optimisation of Administration Route

Formulations as described in Example 4 could also be prepared usingsuitable excipients as intramammary infusions or topical creams. Thesecould be compared against the effect of the drench formulation asoutlined in Examples 2, 3 and/or 4.

Many modifications may be made to the embodiments and examples describedabove without departing from the spirit and scope of the invention.

It is to be understood that, if any prior art publication is referred toherein, such reference does not constitute an admission that thepublication forms a part of the common general knowledge in the art, inAustralia or any other country.

In the claims which follow and in the preceding description of theinvention, except where the context requires otherwise due to expresslanguage or necessary implication, the word “comprise” or variationssuch as “comprises” or “comprising” is used in an inclusive sense, i.e.to specify the presence of the stated features but not to preclude thepresence or addition of further features in various embodiments of theinvention.

1.-19. (canceled)
 20. A method for treating inflammation and/orinfection in breast or udder tissue comprising administering aformulation to a subject in need thereof, wherein the formulationcomprises a non-neutralised tocol phosphate, a vitamin A compound, and ahydrophobic delivery vehicle.
 21. The method of claim 20 wherein thebreast or udder tissue is a mammary gland.
 22. The method of claim 20wherein the inflammation and/or infection is mastitis.
 23. The method ofclaim 20 wherein the subject is selected from humans, domestic mammalsincluding companion animals, working animals, livestock, andzoological/park mammals.
 24. (canceled)
 25. (canceled)
 26. A method forreducing somatic cell count in a lactating subject comprisingadministering a formulation to the lactating subject, wherein theformulation comprises a non-neutralised tocol phosphate, a vitamin Acompound, and a hydrophobic delivery vehicle.
 27. (canceled) 28.(canceled)
 29. A method for supplementing vitamin E levels (μmol/L) in asubject comprising administering a formulation to the subject, whereinthe formulation comprises a non-neutralised tocol phosphate, a vitamin Acompound, and a hydrophobic delivery vehicle.
 30. (canceled) 31.(canceled)
 32. The method of claim 20 wherein the non-neutralised tocolphosphate is selected from the group consisting of non-neutralisedmono-(tocopheryl) phosphate, non-neutralised mono-(tocopheryl) phosphatemonosodium salt, non-neutralised mono-(tocopheryl) phosphate disodiumsalt, non-neutralised mono-(tocopheryl) phosphate monopotassium salt,non-neutralised mono-(tocopheryl) phosphate dipotassium salt,non-neutralised di-(tocopheryl) phosphate, non-neutraliseddi-(tocopheryl) phosphate monosodium salt, non-neutraliseddi-(tocopheryl) phosphate monopotassium salt, non-neutralisedmono-(tocotrienyl) phosphate, non-neutralised mono-(tocotrienyl)phosphate monosodium salt, non-neutralised mono-(tocotrienyl) phosphatedisodium salt, non-neutralised mono-(tocopheryl) phosphate monopotassiumsalt, non-neutralised mono-(tocotrienyl) phosphate dipotassium salt,non-neutralised di-(tocotrienyl) phosphate, non-neutraliseddi-(tocotrienyl) phosphate monosodium salt, non-neutraliseddi-(tocotrienyl) phosphate monopotassium salt, or a combination thereof.33. The method of claim 20 wherein the non-neutralised tocol phosphateis a combination of a non-neutralised mono-(tocopheryl) phosphate and anon-neutralised di-(tocopheryl) phosphate.
 34. The method of claim 33wherein the non-neutralised tocol phosphate is a combination of anon-neutralised mono-(tocopheryl) phosphate and a non-neutraliseddi-(tocopheryl) phosphate and a ratio (w/w %) of non-neutralisedmono-(tocopheryl) phosphate to non-neutralised di-(tocopheryl) phosphateis about 4:1 to about 1:4.
 35. The method of claim 20 wherein thenon-neutralised tocol phosphate has a pH of less than about 4, about 2,about 3, about 2 to about 4, or about 2 to about
 3. 36. The method ofclaim 20 wherein the non-neutralised tocol phosphate comprises fromabout 0.01% w/w up to about 40% w/w of the total concentration of thecomposition.
 37. The method of claim 20 wherein the vitamin A compoundis selected from the group consisting of vitamin A, a derivative ofvitamin A, a metabolite of vitamin A, a precursor of vitamin A, orpro-vitamin A, or a combination thereof.
 38. The method of claim 20wherein the vitamin A compound is a carotenoid.
 39. The method of claim38 wherein the carotenoid is selected from the carotene class includingalpha-carotene, beta-carotene, gamma-carotene, beta-cryptoxanthin andlycopene.
 40. The method of claim 20 wherein the ratio (w/w %) ofnon-neutralised tocol phosphate to vitamin A compound is from about0.01:1 to about 100:1.
 41. The method of claim 20 wherein thenon-neutralised tocol phosphate is a combination of a mono-(tocopheryl)phosphate and a di-(tocopheryl) phosphate, and the vitamin A compound isbeta-carotene.
 42. The method of claim 20 wherein the formulationfurther comprises a nutrient compound selected from the group consistingof antioxidants, vitamins, minerals and trace elements.
 43. The methodof claim 42 wherein the nutrient compound is selected from the groupconsisting of coenzyme Q10, ubiquinol, vitamin D compounds, vitamin Ecompounds, vitamin K compounds, folic acid, vitamin B compounds, vitaminC, vitamin P, vitamin F, lutein, zeaxanthin, cysteine, flavonoids,isoflavones, bilberry, ginkgo biloba, grape seed extract,phytonutrients, alpha lipoic acid, bilberry, bioflavinoids, unsaturatedfatty acids, calcium, phosphorus, magnesium, fluorine, phosphorus,sulfur, sodium, potassium, chloride, calcium, iodine, cobalt, copper,iron, manganese, molybdenum, selenium, zinc, chromium, cadmium,fluorine, nickel, silicon, tin, vanadium, niacin, and combinationsthereof.
 44. The method of claim 20 wherein the hydrophobic deliveryvehicle is an oil or a wax.
 45. The method of claim 20 wherein thehydrophobic delivery vehicle is present in an amount of at least about60.0% w/w of the total concentration of the formulation.